Identification of calcium-binding proteins in fish seminal plasma.

Mariola A Dietrich, Joanna Nynca, Błażej Westfalewicz, Halina Karol, Andrzej Ciereszko

Department of Gametes and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Tuwima 10, 10-714 Olsztyn, Poland. m.dietrich@pan.olsztyn.pl

Fish physiology and biochemistry 2011 Sep

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Calcium ions play an important role in the activation of fish sperm movement. The mechanism of their binding in semen is still unknown. The goal of this study was the development of a method for identifying calcium-binding proteins in fish seminal plasma. Two methods of calcium-binding proteins detection were tested with the use of Quin2 and Stains-all dyes. The first method was useful for the identification of calcium-binding proteins of fish seminal plasma. It consisted of proteins separation using SDS-PAGE, transfer on PVDF membrane, incubation with CaCl₂, staining with Quin2 and illumination with UV light to reveal calcium-binding protein bands. Using Quin2 allowed the detection of calcium-binding proteins with low and high molecular weight. Electrophoretic species-specific profiles of calcium-binding proteins were identified in the seminal plasma of carp, whitefish, roach, brook trout, brown trout and rainbow trout. Staining of calcium-binding proteins with Quin2 is a quick and safe method, allowing the identification of calcium-binding proteins in fish semen.

Citation

Mariola A Dietrich, Joanna Nynca, Błażej Westfalewicz, Halina Karol, Andrzej Ciereszko. Identification of calcium-binding proteins in fish seminal plasma. Fish physiology and biochemistry. 2011 Sep;37(3):447-52